What is ASPETIC FILLING

1. ASPETICFILLING
PHARMACEUTICAL VALIDATION
PRESENTED BY
B.VENKATA SRAVANI
Roll No: 621209521006
I/II – M.PHARMACY
PHARMACEUTICAL ANALYSIS
AU COLLEGE OF PHARMACEUTICAL SCIENCES

2. CONTENTS
•Media fill validation
•USFDA guidelines on process validation- A life cycle
approach
•References

3. WHAT IS A MEDIA FILL?
• A “media fill” is performance of an aseptic manufacturing procedure using a
sterile microbiological growth medium in place of the solution whether the
aseptic procedures are adequate to prevent contamination during actual drug
production.
• Media fill validation is cGMP, FDA and ISO regulatory requirement for
validation of aseptic processing.
• Conducted within manipulations normally performed in actual processing.
• e.g. filling and closing operations, operational environment, processing
operations, number of personnel involved etc are conducted under processing
conditions that include worst case considerations

4. • Why the validation of aseptic process is required by pharmaceutical regulations?
• A “sterile product” is defined as “free of viable organisms”
• As it is not practical examine every unit for confirmation of sterility, all efforts are
made to minimise the risk of contamination (finishing, HVAC, pressure differentials,
cleaning procedure, monitoring programme)
• Despite of many measures taken, contamination is an ever- present danger because
aseptic processing is a process being operated in a controlled –but not sterile-
environment and sample numbers are too small; so that only gross contamination is
likely to be detected

5. • Although media fills must duplicate aseptic manufacturing conditions, it is not
possible for them to be conducted in exactly the same way as the manufacture of a
production batch of a pharmaceutical product.
• In aseptic processing, the greatest risk comes from the personnel working in the clean
room: the operators have to participate in media fills.
• Environmental monitoring activities are required during aseptic filling operations.
• It is usual to include the “worst case” conditions that can occur in production runs.

8. MEDIA FILL PROTOCOL
• Number and frequency of runs
• Medium culture (to replace the product)
• Number of units filled
• Container (vial) size
• Fill volume
• Line speed (or filling speed)
• Duration of fill
• Operators shifts
•Monitoring activities
•Interventions –both routine and non-routine
•Incubation method
•Acceptance criteria

9. NUMBER AND FREQUENCY OF RUNS
• In start up simulation at least three consecutive separate successful runs should be
performed (it is recommended they are performed in different days).
• For on-going simulation, a routine semi-annual qualification is recommended (one
run)
• Extraordinary media fill should be performed after all changes to a product or line
changes evaluated as a potential danger for the aseptic process.

10. MEDIUM CULTURE
• The medium needs to support the growth of a wide variety of microorganisms,
including aerobic bacteria, yeasts and moulds (non-selective medium).
• For aerobic conditions: Soybean Casein Digest Medium (SCD) also known as
tryptone soya broth (TSB).
• For anaerobic conditions: usually in a nitrogen environment fluid thio-glycollate
medium

11. • The media have to support the growth of microorganisms (growth promotion
test).
• The organisms to be tested are stated by pharmacopoeia.
• Generally at the end of incubation period, some vials (taken from the
beginning, at half and at the end of the process) are inoculated with < 100 CFU
and incubated for 3 days (bacteria) and 5 days (yeast and mould)

12. NUMBER OF UNITS FILLED
• Number of units filled should reflect the real batch size.
• It is allowed to fill a lowest number of units provided that the number of units filled
is sufficient to reflect the effect of potential operator fatigue and adequately
represents the maximum number of interventions.
• Some regulations suggest the number of units to be filled in consideration of batch
production size.

13. CONTAINER SIZE
• The extremes of size containers should be considered.
• largest container (often filled at the lowest speed because of its large fill volume)
often has the large opening , so the potential for microbial entry from the
environment should be the greatest for that size.
• The smallest container (often filled at the highest speed for its lower fill volume)
represents the greatest handling difficulty; the smaller container are more fragile and
less stable and be more subjected to breakage and jamming in the equipment.

14. • In the initial qualification two runs might be performed using the largest
container and the third run using the smallest container
• In routine evaluation of the line, any container should be included in the
validation program
• Clear containers should be used as a substitute for amber containers to allow
visual detection of microbial growth

15. FILL VOLUME
• The volume of media filled into the containers need not the routine fill volume.
• It should be sufficient to contact the container-closure seal surfaces (when the unit is
inverted and swirled) and sufficient to allow visual detection of microbial growth post
incubation.
• Smaller containers should not be over-filled as sufficient air must be available in the
container headspace to support the growth of aerobic organisms (generally 25% of
volume is not filled).

16. LINE (OR FILLING) SPEED
• The media fill should address the range of line speeds employed during
production. Sometimes more than one line speed should be evaluated.
• The speed chosen for each batch during simulation should be justified.
• Use of high line speed is justified for manufacturing processes characterized by
frequent interventions or a significant degree of manual manipulation.
• Use of low speed is justified for manufacturing processes characterized by
prolonged exposure of sterile components in the aseptic area.

17. DURATION OF FILL
• In general media fills should be long enough to include all of the required
interventions and stoppage and should reflect the potential operator fatigue: a typical
media fill might be at least 3-4 hours long. Ideally a media fill should use more units
than are in the product being simulated (for all batches up to 5000 units).
• For very large batches or long campaigns, some blank units (either empty or water
filled) are used to maintain operating conditions during the simulation: this
technique can be used to validate processes that may run for several days in order to
validate the full length of the longest approved campaign

18. OPERATORS SHIFTS
• Each operator performing aseptic processes are requested to participate in media
fill.
• Set-up and line operators should be part of not less than one process simulation
per year.
• Operators such as line mechanics and environmental samplers should be managed
in a similar manner.
• A maximum number of personnel present in the aseptic processing room should
be established.

19. • When a firm operates on multiple shifts, the second and third shift should be
included in the media fill programme.
• In case of manual operations (filling), each line operator should participate
into all three initial validation runs and at least one run in re-validation (every
six months)

20. ENVIRONMENTAL MONITORING ACTIVITIES
• There are regulatory and pharmacopoeia references that states the microbial
conditions.
• Air sampling using either active and passive sampling methods should be performed
during the execution of the process. Surface sampling is best performed at the end of
aseptic process. Also personnel should be monitored
• Microbiological monitoring (air, surfaces, personnel) and particle monitoring should
be performed during media fill employing the same procedure in force
• Sometimes the number of sampling locations might be increased respect the routine
procedure

21. INTERVENTIONS –BOTH ROUTINE AND NON-
ROUTINE
• Media fill records should document all interventions performed and the number of
units removed.
• Routine interventions: aseptic line set-up in which sterilised parts are removed from
protective materials and installed is a potential danger; it is common to identify the
first containers filled as they may be more indicative of potential problem with
aseptic assembly.

22. •Other routine interventions: stoppers bowl feeding, remove fallen vials, remove jam
stoppers, operators breaks, gloves change, environmental monitoring.
•Non routine interventions (occur randomly): glass breakage, change / reset of filling
needles,
interventions on weight adjustments, sensor failure, rail adjustments.

23. Incubation methods
• Any filled units should be inspected prior to incubation; any defects that compromise the
container closure or non-integral units are rejected and documented.
• Divergence in industry practice: incubation is performed for 14 days at 20-35°C (+/-
2,5°C): it is performed for 7 days at 20-25°C and further 7 days at 30-35°C; it is
performed for 7 days at 30-35°C and then move the filled containers to 20-25°C
• The lack of agreement suggest that the selection of incubation conditions employed.
• Units are incubated in an inverted position for the first half of the incubation period and
then returned to an upright position for the remainder.

24. Acceptance criteria
• The target should be zero growth but a contamination rate less than 0,10% with
95% confidence level is acceptable (approx. 1 contaminated unit in 5000 filled
units).
• FDA and PDA agree that the target should be zero contaminated units regardless
of size of run
• It is important to note that “invalidation” of a media fill run should be a rare
occurrence
• Each failure should be investigated.

25. •The table indicates the maximum permitted number of contaminated units per
various Media Fill “run sizes” to indicate a 0,10% contamination limit with a 95%
confidence level.
Filled units per run Contaminated units
permitted (action level)
3000
4750
6300
7750
9150
10510
11840
13150
14430
15710
16960
0
1
2
3
4
5
6
7
8
9
10

26. Contamination
• The root cause of a failure (contamination), or at least the most probable one, must be
identified
• It is important to be able to isolate and identify (to species level) the microorganisms
• An appropriate corrective action / preventive action plan must be implemented
• The impact of the failure on product lots already released (if any) must be evaluated
• After the corrective actions have been implemented, a new media fill study is
performed to confirm their efficacy.

28. VALIDATION

29. OBJECTIVE
• The Objective of validation protocol is to establish documented evidence that the
process employed for aseptic processing of Parenterals liquid/Ophthalmic solution
will produce the desired results consistently, within the specified acceptance limits,
when performed as per the latest Standard Operating Procedures.
SCOPE
• The Validation protocol describes the procedure for the total Process Simulation
(Media Fill)

30. Sr. no
.
Responsibility Name of the
department
1. Preparation of Protocol QC
2. Provision of qualified personnel to
assist in the protocol preparation and
execution
QC, QA, Production
and Maintenance
3. Verification of Protocol QC and Production
4. Approval of protocol QA
5. Final determination of System
Acceptability
QA
6. Review and assembling of data into a
final report
QA
RESPONSIBILITIES

31. PRE-REQUISITES
• Approved Soybean casein digest broth
• Environmental Monitoring of manufacturing areas by Plate Exposure, Air sampling
and surface monitoring procedures and its SOP’s.
• Qualified and validated manufacturing equipments, system facility (i.e. HVAC,
water, compressed gases) CIP and SIP procedures.
• Trained operating personnel’s.
• Approved BMR for media fill trial.

32. IDENTIFICATION OF CRITICAL CONTROL MONITORING PARAMETER
• Check and ensure that-
The equipment and system facility is validated.
The HVAC system, compressed air, CIP and SIP procedures are qualified.
All operations, cleaning/sanitization procedures are established and operating
personnel are trained.
Media used for Process Simulation is passed for GPT.
The WFI used for preparation of batch is complied to USP/IP.

33. VALIDATION PROCEDURE
Main steps for the Validation of the integrated line by media fill test
1. Cleaning of the line
2. Dispensing of Soybean Casein Digest Medium for 150 L batch size
3. Batch Preparation 150 L
4. Filling And Sealing
5. Incubation and Examination of Media Filled Units
6. Interpretation of Results

34. VALIDATION PROCEDURE
1. Cleaning of the SVP line
• Carry out cleaning of SVP mixing tank and holding tank along with product line
and bottle pack machine as per respective SOP for CIP.
• At the end of cleaning, collect last rinses sample from sampling point and send to
QC department with written information for testing of previous product traces.
• After getting approval report from QC, affix status label on the tank “READY
FOR STERILIZATION”.
• Immediately carry out the sterilization of SVP holding tank along with final filter
and product line of bottle pack machine as per respective SOP.

35. 2. Dispensing of Soybean Casein Digest Medium for 150 L batch size
• Enter to dispensing room as per SOP for entry exit procedure to dispensing area.
• Check for the clearance of the area from any unwanted materials. Check for the
cleanliness of the area, LAF, weighing pan as per checklist. Put “ON” the reverse
LAF unit 15 minutes before dispensing of material.
• Check the availability of clean containers, pressure differentials, and temperature &
humidity should be not more than 25º C and 45 to 60% RH respectively.

36. • Calibrate the balance as per SOP of Balance Calibration.
• Take the Approved Soybean Casein Digest Medium in pre- dispensing room, place on
SS pallet and check the label of container for correctness and Approval of material.
• Transfer the material to Dispensing room, place the empty clean container on the
balance and record the tare weight. Press “ZERO” of the balance and weigh the
required quantity of material, note the weighed material and then remove the container
from balance and press Zero.
• Close the dispensed material, affix the weighing tag and transfer the material in
dispensed material storage room.

37. • After dispensing, put “OFF” the balance and LAF. Clean the surrounding area,
balance and spray with 70% IPA solution.
• Reseal the original container and shift to their original place.
Batch Preparation 150 L:
• Ensure that the area and product line is clean and free from the traces of previous
product.
• Recheck gross weight of Soybean Casein Digest Medium (SCDM) to be used for
manufacturing and ensure that they match as per entries made in the BMR weighing
sheet.

38. • Check the status board affixed on the tank “READY FOR USE”, also verify the
records and ensure that the bottom outlet valve of the mixing tank is closed.
• Send the entry point sample of WFI from the user point to QC department for
testing along with BMR.
• On approval of WFI sample from QC department, affix a status board on the
Mixing tank “UNDER MANUFCTURING” with Product name and B. No.
• Collect approx 50 L water for injection at 80 to 85º C in a manufacturing tank fitted
with stirrer.
• Start the stirrer and add SCDM through the mainhole of the tank.

39. • Continue stirrer for complete dissolution of ingredients.
• Stop the stirrer.
• Make up the volume to the 150 L with water for injection.
• Start the stirring for complete dissolution of SCDM and homogeneous bulk solution
(generally required 10 minutes).
• Collect sample of bulk solution in a sterile sampling bottle and send it to QC for
testing of color clarity, pH and bioburden along with bulk intimation slip.
• After getting clearance of bulk analysis from Quality Control, start the filtration from
mixing tank to Holding tank of line with the help of pump.

40. • After getting clearance of bulk analysis from Quality Control, start the filtration from
mixing tank to Holding tank of line with the help of pump.
• Perform the bubble point test of the final filter after holding tank as per SOP of Bubble
point test.
4.Filling And Sealing:
• Start the filtration from holding tank to FFS machine using pump.
• Drain one buffer tank approx 1.3 liters of bulk solution from filling nozzle to eliminate
any possibility of dilution of bulk by condensates in product line of the machine post
SIP.

41. • Check online cartridge filter integrity test as per its respective SOP.
• Start Machine line and discard initial 15 shots.
• Collect first cassette of vials from next shot and send the sample with written
information to QC for testing.
• Arrange the out coming cassettes of vials sequentially in vacuum chamber tray and
verify the results of testing from QC department.
• Now start the filling and sealing continuously as per SOP for Filling and sealing.

42. • Collect the filled and sealed containers coming out of the filling area in plastic crates.
• During filling operation keep the filled ampoules separately for each breakdown,
shift change, power breakdown, stoppage etc and assign lot number.
• Arrange the cassettes of vials lot wise in SS trays vertically in vacuum leak testing
chamber tray and carry out the leak testing at 650 – 720 mm Hg for 30 minutes. Do
not use the leak vials for further media fill study.
• After leak test, transfer the goods vials in the clean plastic crates horizontally in
cassette from one above the other, lot wise separately

43. 5. Incubation and examination of filled units:
a. Incubate all media filled units in normal position after leak test at of 20 to 250 C for 7
days. Incubation temperature should be maintained within 22.5 ± 2.50 C .
b. After completion of 7 days Incubation at 20 to 250 C, invert the units and incubate them
at 30-350 C for next 7 days. Incubation temperature should be maintained within
32.5±2.50 C
c. Each media filled unit should be examined by trained Microbiologist after 3rd day, 7th
day, 10th day and 14th day.
d. All suspect units identified during the observation should be brought to the immediate
attention of the QC Microbiologist.

44. Interpretation of Results:
• When filling fewer than 5,000 units, no contaminated units should be detected.
When filling 5,000 to 10,000 units :
• One contaminated unit should result in an investigation, including consideration of a
repeat media fill
• Two contaminated units are considered cause for revalidation, following investigation.
When filling more than 10,000 units :
• One contaminated unit should result in an investigation;
• Two contaminated units are considered cause for revalidation, following investigation

45. USFDA GUIDELINES ON
PROCESS VALIDATION –
LIFECYCLE APPROACH

46. Introduction
• Validation of manufacturing processes is a requirement of the Current Good
Manufacturing Practice (CGMP) regulations for finished pharmaceuticals (21 CFR
211.100 and 211.110) and is considered as an enforceable element of Current Good
Manufacturing Practice for active pharmaceutical ingredients (APIs) under the broader
statutory CGMP provisions of section 501(a)(2)(B) of the Federal Food, Drug and
Cosmetic Act.
• Process validation involves a series of activities taking place over the lifecycle of the
product and process.”

47. Definition of Process Validation
• Stated in the 2011 guidance is as follows:
“Process validation is defined as the collection and evaluation of data, from the process
design stage throughout production, which establishes scientific evidence that a process is
capable of consistently delivering quality product”
• 1987 Definition :
“Establishing documented evidence which provides a high degree of assurance that a
specific process will consistently produce a product meeting its predetermined
specifications and quality characteristics”

48. Approach to Process Validation
• Stage 1: Process Design: The marketable manufacturing process is defined during this
stage based on knowledge gained through development and scale-up activities.
• Stage 2: Process Qualification: Throughout this stage, the method design is estimated
to determine if the process is capable of reproducible marketable business.
• Stage 3: Continued Process Verification: Constant assertion is gained during routine
production that the process remains in a state of control.

49. Stage1: Process Design:
Constructing and Apprehending Process Knowledge and Understanding:
• The functionality and limits of commercial manufacturing equipment should be
considered in the process design.
• Design of experiments (DOE) studies can help to develop process knowledge by
revealing relationships, including multivariate interactions, between the variable inputs
and the resulting outputs.

50. Stage 2: Process Qualification:
Element (1): Design of a facility and qualification of utilities and equipment
• Ensure qualification of facility, utilities and equipment is completed & documented prior
to initiate process qualification.
Element (2): Process Performance Qualification (PPQ)
• The PPQ combines the actual facility, utilities, equipment’s and the trained personnel
with the commercial manufacturing controls.
• A company must successfully complete PPQ before commencing commercial
distribution of the drug product.

51. • Strongly recommend firms employ objective measure (e.g. Statistical Metrics) wherever
feasible and meaningful to achieve adequate assurance.
• The increased level of inspection, testing, and sampling should continue through the
process verification stage as correct, to establish levels and occurrence of routine
sampling and checking for the particular product and process.
• Considerations for the duration of the intensified sampling & checking period could
include (not limited to):
Volume of production
Process Complexity
Level of process understanding
Experience with similar products and process.

52. PPQ Protocol:
• A written protocol that specifies the manufacturing conditions, controls, testing, and
expected outcomes is essential for this stage of process validation.
• Protocol discuss the following elements:
• The manufacturing conditions, including operating parameters, processing limits, and
component (raw material) inputs.
• The data to be collected and when and how it will be evaluated.
• Tests to be performed (in-process, release, characterization) and acceptance criteria for
each significant processing step.
52

53. • The sampling plan, including sampling points, number of samples, and the frequency
of sampling for each unit operation and attribute.
• The number of samples should be adequate to provide sufficient statistical confidence
of quality both within a batch and between batches.
• The confidence level selected can be based on risk analysis as it relates to the particular
attribute under examination. Sampling during this stage should be more extensive than
is typical during routine production.

54. • Criteria and process performance indicators that allow for a science- and risk-based
decision about the ability of the process to consistently produce quality products. The
criteria should include:
1. A description of the statistical methods to be used in analyzing all collected data (e.g.,
statistical metrics defining both intra-batch and inter-batch variability).
2. Provision for addressing deviations from expected conditions and handling of
nonconforming data.
Data should not be excluded from further consideration in terms of PPQ
without a documented, science-based justification.

55. • Design of facilities and the qualification of utilities and equipment, personnel
training and qualification, and verification of material sources (components and
container/closures), if not previously accomplished.
• Status of the validation of analytical methods used in measuring the process, in-
process materials, and the product.
• Review and approval of the protocol by appropriate departments and the quality
unit.

56. PPQ Report:
• To state a clear conclusion as to whether the data indicates the process meets the
conditions established in the protocol. If not the report should state what should be
accomplished before such a conclusion can be reached.
• This conclusion should be based on entire compilation of knowledge and information
gained from the design stage through the PPQ stage.

57. Stage 3: Continued Process Verification:
• To confirm that “the process remains in a state of control during commercial
manufacture.” An ongoing process to collect and analyse product and process data
that relate to product quality must be established.
• The results obtained should be statistically trended and reviewed by trained
personnel. Recommend that a person with suitable training in statistical process
control techniques develop the data collection plan and statistical methods.
• Good process design and development should anticipate significant sources of
variability and establish appropriate detection, control and or qualification schemes,
as well as suitable alert and action limits.

58. • Study of intra-batch as well as inter-batch variation is part of a comprehensive
continued process verification program.
• Deviation can be detected by the timely assessment of
1. Defect complaints,
2. OOS findings,
3. Process deviation report,
4. Process yield variations,
5. Batch record & reports
• Manufacture line operatives and quality unit staff should be encouraged to provide
feedback on process performance.

59. • Quality unit meet periodically with production staff to evaluate data, discuss possible
trends and coordinate any correction or follow-up actions by product.
• Data collected during this stage might recommend ways to improve and/or optimize
the process by altering some aspect of the process or product, such as the operating
conditions, process controls, etc.
• Well justified rationale for the change, implementation plan, quality unit approval
before implementation.

60. Documentation
• Documentation at each stage of the process validation lifecycle is essential for effective
statement in difficult, lengthy, and multidisciplinary projects.
• Documents is important so that knowledge gained about a product and process is
accessible and comprehensible to others involved in each stage of the lifecycle.
• The degree and type of records required by CGMP vary during the validation lifecycle.
Records requirements are greatest during Stage 2, process requirement, and Stage 3,
continued process confirmation. Studies during these stages must conform to CGMPs
and must be approved by the quality unit in accordance with the regulations.
60

61. WHY THE LIFECYCLE APPROACH?
• For manufacturing processes to be truly validated, each of the stages must be
addressed and integrated. This integration of development work, process conformance,
and continuing verification provides assurance that the product or process will
consistently remain in control throughout the entire product lifecycle.
• Process validation must not be considered a one-time event or a focused one-time task
performed just prior to commercial launch that emphasizes only the manufacture of
three conformance lots.
• Acceptable manufacture of three conformance batches must not be interpreted as
completion of validation.

62. • These lots cannot truly represent the future manufacturing process with unexpected
and unpredictable changes.
• Conformance lots are often inadvertently biased (i.e., they may utilize well-
characterized and controlled API and excipients, be manufactured under well
controlled conditions, be monitored by expert individuals, and performed by most
experienced or well- trained personnel—all “best-case” conditions).
• It is highly unrealistic to contend that the manufacture of three conformance lots under
“best-case” conditions conclusively predicts successful manufacturing over the
product lifetime. True process validation must be a process that is never completed
and is always ongoing.

63. Positive Aspects Of Life Cycle Approach:
• New life cycle validation model is a science and risk based approach and consistent
with the “Quality by Design” approach that is articulated in ICH guidelines Q-8, Q-9,
Q- 10 & Q11, positive aspects of the new approach are as follows:
1. Robust process validation approach will lead to consistent and reliable quality of
product.
2. Reduction in cost of quality, time and energy.
3. Ongoing statistical evaluation of data will detect the trending at early stage to avoid
potential failures at later stages.

64. 4. Right first time solutions can be offered in avoiding reduction in rejections/recycles.
5. Continuous improvement overtime rather than incidental based such as
failures/OOS.
6. Increased into the machine efficiency in enhancing the productivity outlet.
7. Implementation of real time release testing in lieu of end product testing based on
statistical evaluation.
8. Relief in drug application approval process and reduction/fast approval in post
approval changes.

65. CONCLUSION
• By doing validation as per USFDA process validation guideline, product and process
understanding will be improved and also reduction in waste, rejections, lead time and
any other failures.
• This guideline also helps for continual improvement of validation process through the
product life cycle.

66. REFERENCES
• R. A. Nash and A. H. Wachter “Pharmaceutical Process validation”; Third edition
• Agallo James, Carleton J. Fredric “Validation of Pharmaceutical Processes”; Third edition
• USP <797> ‘media fill testing’ / <71> ‘growth promotion test’
• FDA “guidance for industry, sterile drug products produced by aseptic processing – cGMP”
6. PIC/S PI 007-2 “recommendations on the validation of aseptic process on 28/02/2020
• https://www.fda.gov/media/81974/download on dated 07/02/2020
• FDA (CDER and CBER), Process Validation: General Principles and Practices, Current
Good Manufacturing Practices (CGMP), Revision 1,January 2011.

67. Thank You